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      The Metabolomic Profile of Spent Culture Media from Day-3 Human Embryos Cultured under Low Oxygen Tension

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          Despite efforts made to improve the in vitro embryo culture conditions used during assisted reproduction procedures, human embryos must adapt to different in vitro oxygen concentrations and the new metabolic milieu provided by the diverse culture media used for such protocols. It has been shown that the embryo culture environment can affect not only cellular metabolism, but also gene expression in different species of mammalian embryos. Therefore we wanted to compare the metabolic footprint left by human cleavage-stage embryos under two types of oxygen atmospheric culture conditions (6% and 20% O 2). The spent culture media from 39 transferred and implanted embryos from a total of 22 patients undergoing egg donation treatment was analyzed; 23 embryos came from 13 patients in the 6% oxygen concentration group, and 16 embryos from 9 patients were used in the 20% oxygen concentration group. The multivariate statistics we used in our analysis showed that human cleavage-stage embryos grown under both types of oxygen concentration left a similar metabolic fingerprint. We failed to observe any change in the net depletion or release of relevant analytes, such as glucose and especially fatty acids, by human cleavage-stage embryos under either type of culture condition. Therefore it seems that low oxygen tension during embryo culture does not alter the global metabolism of human cleavage-stage embryos.

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          Most cited references 57

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          'Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data.

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            Visualization of GC/TOF-MS-based metabolomics data for identification of biochemically interesting compounds using OPLS class models.

            Metabolomics studies generate increasingly complex data tables, which are hard to summarize and visualize without appropriate tools. The use of chemometrics tools, e.g., principal component analysis (PCA), partial least-squares to latent structures (PLS), and orthogonal PLS (OPLS), is therefore of great importance as these include efficient, validated, and robust methods for modeling information-rich chemical and biological data. Here the S-plot is proposed as a tool for visualization and interpretation of multivariate classification models, e.g., OPLS discriminate analysis, having two or more classes. The S-plot visualizes both the covariance and correlation between the metabolites and the modeled class designation. Thereby the S-plot helps identifying statistically significant and potentially biochemically significant metabolites, based both on contributions to the model and their reliability. An extension of the S-plot, the SUS-plot (shared and unique structure), is applied to compare the outcome of multiple classification models compared to a common reference, e.g., control. The used example is a gas chromatography coupled mass spectroscopy based metabolomics study in plant biology where two different transgenic poplar lines are compared to wild type. By using OPLS, an improved visualization and discrimination of interesting metabolites could be demonstrated.
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              Oxygen tension in the oviduct and uterus of rhesus monkeys, hamsters and rabbits.

              Oxygen tension was measured using flexible polarographic microelectrodes within the oviductal and uterine lumen in rhesus monkeys (n = 9), golden hamsters (n = 21) and rabbits (n = 6), during the reproductive cycle (monkey), during oestrus and pseudopregnancy (hamsters, rabbits) and during pregnancy (hamsters). In general, oxygen tensions in each species were much less than half of atmospheric O2, ranging from high values of about 60 mm Hg (8.7% O2) in the rabbit oviduct, rabbit and hamster uterus, to as low as 11 mm Hg (1.5% O2) in the monkey uterus. Oxygen tensions did not vary significantly between left and right sides of the reproductive tracts (all species), nor between pregnant and pseudopregnant states nor between oviduct and uterus (hamsters). Differences owing to reproductive stage were found in the monkey oviduct, hamster oviduct and uterus, and rabbit uterus. Oxygen tensions were consistently very low (11-14 mm Hg) in the monkey uterus throughout the menstrual cycle. In hamsters and rabbits, intrauterine O2 decreased significantly at about the normal time of blastocyst formation and implantation, to 37 mm Hg (5.3% O2) and 24 mm Hg (3.5% O2), respectively. This study indicates that embryos develop in vivo under low oxygen concentrations, especially during the peri-implantation period. The data have implications for investigations of embryo metabolism and for improving embryo development in vitro.

                Author and article information

                [1 ]IVI Valencia, Valencia, Spain
                [2 ]INCLIVA Biomedical Research and Fundación IVI, Valencia, Spain
                [3 ]IVI Sevilla, Sevilla, Spain
                [4 ]OWL, Derio, Spain
                Peiking university third hospital, CHINA
                Author notes

                Competing Interests: José Remohí is employee of IVI Valencia and also stockholder of the company. Maria José de los Santos, Pilar Gámiz and Josep Lluís Romero are employees of IVI Valencia. Nicolás Prados is employee of IVI Sevilla. Cristina Alonso is employee of OWL (trading name of One Way Liver Genomics, S.L.). Francisco Dominguez have no conflicts of interest to declare. This does not alter the authors’ adherence to all the PLoS ONE policies on sharing data and materials.

                Conceived and designed the experiments: MJDLS FD. Performed the experiments: MJDLS JMDLS PG NP JR. Analyzed the data: CA FD. Contributed reagents/materials/analysis tools: PG JMDLS JR NP. Wrote the paper: MJDLS JR FD.

                Role: Editor
                PLoS One
                PLoS ONE
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                12 November 2015
                : 10
                : 11
                26562014 4643011 10.1371/journal.pone.0142724 PONE-D-15-21736

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                Figures: 4, Tables: 2, Pages: 15
                FD’s participation was supported by the Spanish Ministry of Economy and Competitiveness, through the Miguel Servet Programme (CP13/00075) co-founded by FEDER and by the R+D programme from the Generalitat Valenciana (Regional Valencian Government) IMPIVA MIDTF/2011/214. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Research Article
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                All relevant data are within the paper and its Supporting Information files.



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